Genetic typing of classical swine fever virus

Three regions of the classical swine fever virus (CSFV) genome that have been widely sequenced were compared with respect to their ability to discriminate between isolates and to segregate viruses into genetic groups. Sequence data-sets were assembled for 55 CSFVs comprising 150 nucleotides of the 5' non-translated region, 190 nucleotides of the E2 envelope glycoprotein gene and 409 nucleotides of the NS5B polymerase gene. Phylogenetic analysis of each data-set revealed similar groups and subgroups. For closely related viruses, the more variable or larger data-sets gave better discrimination, and the most reliable classification was obtained with sequence data from the NS5B region. No evidence was found for intertypic recombination between CSFVs. A larger data-set was also analysed comprising 190 nucleotides of E2 sequence from 100 CSFVs from different parts of the world, in order to assess the extent and global distribution of CSFV diversity. Additional groups of CSFV are evident from Asia and the nomenclature of Lowings et al. (1996) [Lowings, P., Ibata, G., Needham, J., Paton, D., 1996. J. Gen. Virol. 77, 1311-1321] needs to be updated to accommodate these. A tentative assignment, adapting rather than overturning the previous nomenclature divides CSF viruses into three groups with three or four subgroups: 1.1, 1.2, 1.3; 2.1, 2.2, 2.3; 3.1, 3.2, 3.3, 3.4. The expanding data-base of CSFV sequences should improve the prospects of disease tracing in the future, and provide a basis for a standardised approach to ensure that results from different laboratories are comparable.     

Effects of supplemental protein type on productivity of primiparous beef cows

Effects of supplemental degradable (DIP) and undegradable (UIP) intake protein on forage intake, BW change, body condition score (BCS), postpartum interval to first estrus, conception rate, milk production and composition, serum metabolites and metabolic hormones, and calf gain were determined using 36 primiparous Gelbvieh x Angus rotationally crossed beef cows. On d 3 postpartum, cows (average initial BW = 495 +/- 10 kg and BCS = 5.5 +/- 0.1) were randomly assigned to one of three dietary supplements (12 cows/treatment). Date of parturition was evenly distributed across treatment (average span of calving date among treatments = 2.4 +/- 2.5 d). Individually fed (d 3 through 120 postpartum) dietary supplements were 0.82 kg of corn and 0.23 kg of soybean meal per day (DIP), the DIP + 0.12 kg of blood meal and 0.13 kg of corn gluten meal per day (DIP + UIP), and 0.82 kg of corn, 0.07 kg of blood meal, and 0.08 kg of corn gluten meal per day in an isonitrogenous replacement of soybean meal (UIP IsoN). Cows had ad libitum access to native grass hay (8.5% CP) and trace-mineralized salt. Total OM intake was greater (P = 0.06) for DIP + UIP than UIP IsoN cows. At 30 d postpartum, DIP + UIP cows produced more milk than UIP IsoN, with DIP being intermediate; however, at 60 d postpartum, DIP + UIP and DIP cows were not different, but both had greater milk production than UIP IsoN (treatment x day interaction; P = 0.08). A treatment x day interaction (P = 0.06) for BCS resulted from DIP + UIP cows having the greatest BCS at 60, 90, and 120 d d postpartum and DIP having greater BCS than UIP IsoN cows only on d 60 postpartum. Serum insulin concentrations were highest (treatment x day interaction; P = 0.09) for DIP + UIP cows at 30 d postpartum but did not differ among treatment thereafter. Serum insulin-like growth factor-binding protein (IGFBP)-2 (34 kDa) and -3 (40 and 44 kDa) were greatest (P < 0.0003) for DIP cows. Serum urea-N concentrations were greater (P < 0.01) in DIP + UIP cows than in either DIP or UIP IsoN cows. However, postpartum interval to first estrus, conception rate, and calf weaning weights were unaffected (P = 0.35, 0.42, and 0.64, respectively) by treatment. Although UIP in addition to or in replacement of DIP affected milk production and blood metabolites, the productivity of these primiparous beef cows was not altered. Thus, the type of supplemental protein does not seem to influence productivity of primiparous beef cows in production systems with conditions similar to our experimental conditions.     

First report of leaf spot caused by <Emphasis Type="Italic">Alternaria alternata</Emphasis> on <Emphasis Type="Italic">Drimia maritima</Emphasis>

Drimia maritima (squill) is a historically important medicinal plant. During the spring of 2016, small, yellow leaf spots, which became brown and finally necrotic, were observed on squill plants in Kohgiluyeh and Boyer-Ahmad Provinces in Iran. A fungus was consistently isolated from infected leaves and identified as Alternaria alternata based on morphological and phylogenetic analyses. Pathogenicity tests confirmed A. alternata to be the causal agent of the newly observed leaf spot disease. This is the first report of leaf spot on D. maritima caused by A. alternata in the world.     

In silico prediction of Gallibacterium anatis pan-immunogens

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0080-0) contains supplementary material, which is available to authorized users.     

Accumulation of Sapogenin Conjugates and Histological Changes in the Liver and Kidneys of Lambs Suffering from Alveld,a Hepatogenous Photosensitization Disease of Sheep Grazing Narthecium ossifragum

Sixteen lambs exhibiting hepatogenous photosensitization (alveld) after grazing pasture containing Narthecium ossifragum and seven nonphotosensitized lambs grazing the same pastures were studied. All the alveld-affected lambs revealed liver damage dominated by single cell necrosis, portal fibroplasia and bile duct proliferation. Crystalloid clefts were demonstrated in the bile ducts of two and in the hepatocytes and Kupffer cells of nine photosensitized lambs. Plasma bilirubin concentration was severely increased in ten of the cases of alveld whereas the activity of aspartate aminotransferase was moderately to severely increased in seven cases. The activity of glutamate dehydrogenase was moderately elevated in one of the photosensitized lambs. The main histopathological findings in the kidneys from the alveld-affected lambs were dilated tubules, often with eosinophilic material in the tubular lumina. Regenerative changes were seen in a large proportion of the renal sections. Elevated plasma concentrations of urea and creatinine, and the renal histopathological changes, suggested that the photosensitized lambs had been through a phase of renal injury. Analysis of the free and conjugated sapogenin content in liver tissue and bile was performed by gas chromatography–mass spectrometry. There were significantly higher concentrations of conjugated episapogenins in both the liver and bile in the alveld-affected lambs than in the nonphotosensitized lambs.     

Cardiopulmonary and sedative effects of intravenous or epidural methadone in conscious dogs

Cardiopulmonary and sedative effects of intravenous or epidural methadone were compared. Six beagles were randomly assigned to group MIV (methadone 0.5 mg/kg IV + NaCl 0.9% epidurally) or MEP (methadone 0.5 mg/kg epidurally + NaCl 0.9% IV). Cardiopulmonary, blood gas and sedation were assessed at time (T) 0, 15, 30, 60, 120, 240 and 480 min after drug administration. Compared to T0, heart rate decreased at T15–T120 in MIV (p < .001) and T15–T240 in MEP (p < .05); mean arterial pressure was reduced at T15–T60 in MEP (p < .01); respiratory rate was higher at T15 and T30 in both groups (p < .05); pH was lower at T15–T120 in MIV (p < .01) and T15, T30 and T120 in MEP (p < .05); PaCO₂ was higher at T15–T60 in MIV (p < .01) and T15, T30 and T120 in MEP (p < .01); sedation scores were higher at T15 and T30 in MIV and T15–T60 in MEP (p < .05). At T120 and T240, sedation score was higher in group MEP compared with group MIV (p < .01) In conclusion, cardiopulmonary and sedative effects of identical methadone doses are similar when administered IV or epidurally to conscious healthy dogs.     

Callose and β‐1,3‐glucanase inhibit Phytophthora cinnamomi in a resistant avocado rootstock

Phytophthora root rot (PRR) of avocado, caused by Phytophthora cinnamomi, is a significant threat to sustainable production wherever the crop is grown. Resistant rootstocks in combination with phosphite applications are the most effective options for managing this disease. Recently, the mechanisms underpinning PRR resistance have been investigated by the avocado community. Here, biochemical assays and confocal and scanning electron microscopy were used to investigate early defence responses in PRR resistant and ‐susceptible avocado rootstocks. Zoospore germination and subsequent hyphal growth for the pathogen were significantly inhibited on the surface of resistant avocado roots. When penetration occurred in the resistant R0.06 rootstock, callose was deposited in the epidermal cells, parenchyma and cortex of roots. In addition, β‐1,3‐glucanase was released early (6 h post‐inoculation, hpi) in response to the pathogen, followed by a significant increase in catalase by 24 hpi. In contrast, susceptible R0.12 roots responded only with the deposition of lignin and phenolic compounds incapable of impeding pathogen colonization. In this study, PRR resistance was attributed to a timely multilayered response to infection by P. cinnamomi.     

Effect of tillage depth on physical properties of a tropical soil and on yield of maize, sorghum and cotton

Field experiments were carried out to study the effect of three tillage depths (5, 15 and 30 cm) on soil physical properties and on yield of maize (Zea mays), sorghum (Sorghum bicolor) and cotton (Gossypium hirsutum) on a ferruginous tropical soil. The increase in porosity due to deep tillage was only temporary and differences in water storage and movement were only noticeable during the early part of the rainy season. Deep tillage increased the yield of maize and cotton by about 10% but sorghum yield was not affected.